RU2134267C1 - Rapamycin hydroxyesters, method of their synthesis and methods of treatment - Google Patents

Abstract

FIELD: organic chemistry, pharmacy. SUBSTANCE: invention describes new compounds of the formula (I) where each of R¹ and R² means independently hydrogen atom or -CO(CR³R⁴)_b(CR⁵R⁶)_dCR⁷R⁸R⁹ where each of R³ and R⁴ means independently hydrogen atom, alkyl with 1-6 carbon atoms; each of R⁵ and R⁶ means independently hydrogen atom, alkyl with 1-6 carbon atoms or R⁵ and R⁶ can form in common the group X or cycloalkyl ring with 3-8 carbon atoms; R⁷ means hydrogen atom, alkyl with 1-6 carbon atoms; each of R⁸ and R⁹ means independently hydrogen atom, alkyl with 1-6 carbon atoms, -(CR³R⁴)_fOR¹⁰ or R⁸ and R⁹ can in common form groups X or cycloalkyl ring with 3-8 carbon atoms; R¹⁰ means hydrogen atom, alkyl with 1-6 carbon atoms, tri (alkyl) with 1-6 carbon atoms silyl, tri (alkyl) with 1-6 carbon atoms silylethyl or tetrahydropyranyl; X means 5-[(2,2-di(alkyl] c 1-6 carbon atoms (1,3)-dioxanyl, 5-(2-spirocycloalkyl) with 3-8 carbon atoms (1,3)-dioxanyl, 4-[2,2-di(alkyl)] with 1-6 carbon atoms (1,3)-dioxanyl, 4-[2-spiro(cycloalkyl)] with 3-8 carbon atoms (1,3)-dioxanyl; b = 0-6; d = 0-6; f = 0-6 at condition that R¹ and R² are not hydrogen atom simultaneously and either R¹ or R² has at least one (CR³R⁴)_fOR¹⁰, X or cycloalkyl with 3-8 carbon atoms. Pharmaceutically acceptable salt of this compound is used as an immunosuppressive, anti-inflammatory, antifungal, antiproliferative and antitumor agent. Invention describes also a method of synthesis of compounds of the formula (I) showing immunosuppressive activity and a method of treatment of patients using compounds of the formula (I). EFFECT: improved method of synthesis, enhanced effectiveness of compounds. 22 cl, 14 ex

Description

 The invention relates to hydroxyesters, to a method of using such compounds for inducing immunosuppression, as well as for treating rejection during transplantation or incompatibility reactions of autoimmune diseases, inflammatory diseases, adult T-cell leukemia / lymphoma, solid tumors, fungal infections and disorders associated with vascular hyperproliferation .

 Rapamycin is a macrocyclic triene antibiotic produced by Streptomyces hygroscopicus; rapamycin was found to have an antifungal effect, especially against Candida albicans, both in vivo and in vitro. (C. Vezina et al., J. Antibiot. 28, 721 (1975); SN Sehgal et al., J. Antibiot 28,727 (1975); HA Baker et al., J. Antibiot. 31, 539 (1978); U.S. Patent 3,929,992; and U.S. Patent 3,993,749).

 It has been found that rapamycin (US Pat. No. 4,885,171) or in combination with picybanil (US Pat. No. 4,401,653) has antitumor activity. R. Martel et al., (Can. J.Physiol. Pharmacol. 55, 48 (1977)) indicate that rapamycin demonstrated efficacy in an experimental model of allergic encephalomyelitis, in models of multiple sclerosis, adjuvant arthritis, rheumatoid arthritis, and while suppressing the formation of antibodies similar to IgE.

 The immunosuppressive effect of rapamycin is described in FASEB 3, 3411 (1989). It was found that cyclosporin A and FK-506, other macrocyclic molecules, are also effective as immunosuppressive agents, and therefore are used to prevent transplant rejection (FASEB 3, 3411 (1989); FASEB 3, 5256 (1989); RY Calne et al. , Lancet 1183 (1978); and U.S. Patent 5,100,899).

 It has been found that rapamycin can also be used to prevent or treat systemic lupus erythematosus (US Pat. No. 5,078,999), pneumonia (US Pat. No. 5,080,899), diabetes mellitus with insulin dependence (Fifth Int. Conf. Inflamm. Res. Assoc. 121 (abstract), (1990)), smooth muscle cell proliferation and thickening of the inner lining due to vascular damage (Morris, RJ Heart Lung Transplant 11 (pt 2); 197 (1992)), adult T-cell leukemia / lymphoma (Application for Europatent 525960 A1) as well as ocular inflammation (Application for Europatent 532862 A1).

 It has been found that mono- and diacylated derivatives of rapamycin (esterified at 28 and 43 positions) can be used as antifungal agents (US Pat. No. 4,316,885) and are used to prepare water-soluble aminoacyl prodrugs of rapamycin (US Pat. No. 4,650,803). Recently, the numbering rules for rapamycin have been changed; therefore, in accordance with the Nomenclature of Chemical Abstracts, the above esters will be in positions 31 and 42.

где каждый из R¹ и R², независимо, представляет собой водород или
-CO(CR³R⁴)_b (CR⁵R⁶)_d CR⁷R⁸R⁹;
каждый из R³ и R⁴, независимо, представляет собой водород, алкил с 1-6 атомами углерода, алкенил с 2-7 атомами углерода, алкинил с 2-7 атомами углерода, трифторметил или -F;
каждый из R⁵ и R⁶, независимо, представляют собой водород, алкил с 1-6 атомами углерода, алкенил с 2-7 атомами углерода, алкинил с 2-7 атомами углерода, (CR³R⁴)_fOR¹⁰, -CF₃, F, или CO₂R¹¹, или R⁵ и R⁶ могут вместе образовывать X или циклоалкильное кольцо с 3-8 атомами углерода, которое может быть моно-, ди- или тризамещенными -(CR³R⁴)_fOR¹⁰;
R⁷ представляет собой водород, алкил с 1-6 атомами углерода, алкенил с 2-7 атомами углерода, алкинил с 2-7 атомами углерода, (CR³R⁴)_fOR¹⁰, -CF₃, -F, или CO₂R¹¹;
каждый из R⁸ и R⁹, независимо, представляют собой водород, алкил с 1-6 атомами углерода, алкенил с 2-7 атомами углерода, алкинил с 2-7 атомами углерода, -(CR³R⁴)_fOR¹⁰, -CF₃, -F, или R⁸ и R⁹ могут вместе образовывать X или циклоалкильное кольцо с 3-8 атомами углерода, которое может быть моно-, ди- или тризамещенным радикалом -(CR³R⁴)_fOR¹⁰;
R¹⁰ представляет собой водород, алкил с 1-6 атомами углерода, алкенил с 2-7 атомами углерода, алкинил с 2-7 атомами углерода, три(алкил с 1-6 атомами углерода) силил, три(алкил с 1-6 атомами углерода) силилэтил, трифенилметил, бензил, алкоксиметил с 2-7 атомами углерода, три(алкил с 1-6 атомами углерода) силилэтоксиметил, хлорэтил или титрагидропиранил;
R¹¹ представляет собой водород, алкил с 1-6 атомами углерода, алкенил с 2-7 атомами углерода, алкинил с 2-7 атомами углерода, или фенилалкил с 7-10 атомами углерода;
X представляет собой 5-(2, 2-ди(алкил с 1-6 атомами углерода))[1,3]диоксанил, 5-(2-спиро(циклоалкил с 3-8 атомами углерода))[1,3]диоксанил, 4-(2,2-ди(алкил с 1-6 атомами углерода))[1,3]-диоксанил, 4-(2-спиро(циклоалкил с 3-8 атомами углерода))[1,3] -диоксанил, 4-(2,2-ди-(алкил с 1-6 атомами углерода))[1,3]-диоксаланил, или 4-(2-спиро(циклоалкил с 3-8 атомами углерода))[1,3]диоксаланил;
b = 0-6;
d = 0-6;
f = 0-6,
при условии, что R¹ и R² не одновременно являются водородом, а также при условии, что либо R¹, либо R² содержит по меньшей мере один (CR³R⁴)_fOR¹⁰, X или циклоалкил с 3-8 атомами углерода, замещенный (CR³R⁴)_fOR¹⁰ или фармацевтически приемлемую соль этого соединения.The present invention provides derivatives of rapamycin, which are applicable as immunosuppressive, anti-inflammatory, antifungal, antiprofilerative and antitumor agents, and which have the following structure:

where each of R ¹ and R ² independently represents hydrogen or
-CO (CR ³ R ⁴ ) _b (CR ⁵ R ⁶ ) _d CR ⁷ R ⁸ R ⁹ ;
each of R ³ and R ⁴ , independently, is hydrogen, alkyl with 1-6 carbon atoms, alkenyl with 2-7 carbon atoms, alkynyl with 2-7 carbon atoms, trifluoromethyl or -F;
each of R ⁵ and R ⁶ independently represents hydrogen, alkyl with 1-6 carbon atoms, alkenyl with 2-7 carbon atoms, alkynyl with 2-7 carbon atoms, (CR ³ R ⁴ ) _f OR ¹⁰ , -CF ₃ , F, or CO ₂ R ¹¹ , or R ⁵ and R ⁶ may together form an X or cycloalkyl ring with 3-8 carbon atoms, which may be mono-, di- or trisubstituted - (CR ³ R ⁴ ) _f OR ¹⁰ ;
R ⁷ represents hydrogen, alkyl with 1-6 carbon atoms, alkenyl with 2-7 carbon atoms, alkynyl with 2-7 carbon atoms, (CR ³ R ⁴ ) _f OR ¹⁰ , —CF ₃ , —F, or CO ₂ R ¹¹ ;
each of R ⁸ and R ⁹ independently represents hydrogen, alkyl with 1-6 carbon atoms, alkenyl with 2-7 carbon atoms, alkynyl with 2-7 carbon atoms, - (CR ³ R ⁴ ) _f OR ¹⁰ , - CF ₃ , -F, or R ⁸ and R ⁹ may together form an X or cycloalkyl ring with 3-8 carbon atoms, which may be a mono-, di- or trisubstituted radical - (CR ³ R ⁴ ) _f OR ¹⁰ ;
R ¹⁰ represents hydrogen, alkyl with 1-6 carbon atoms, alkenyl with 2-7 carbon atoms, alkynyl with 2-7 carbon atoms, three (alkyl with 1-6 carbon atoms) silyl, three (alkyl with 1-6 atoms carbon) silylethyl, triphenylmethyl, benzyl, alkoxymethyl with 2-7 carbon atoms, three (alkyl with 1-6 carbon atoms) silylethoxymethyl, chloroethyl or titrahydropyranyl;
R ¹¹ represents hydrogen, alkyl with 1-6 carbon atoms, alkenyl with 2-7 carbon atoms, alkynyl with 2-7 carbon atoms, or phenylalkyl with 7-10 carbon atoms;
X represents 5- (2, 2-di (alkyl with 1-6 carbon atoms)) [1,3] dioxanyl, 5- (2-spiro (cycloalkyl with 3-8 carbon atoms)) [1,3] dioxanyl , 4- (2,2-di (alkyl with 1-6 carbon atoms)) [1,3] -dioxanyl, 4- (2-spiro (cycloalkyl with 3-8 carbon atoms)) [1,3] -dioxanyl , 4- (2,2-di- (alkyl with 1-6 carbon atoms)) [1,3] -dioxalanyl, or 4- (2-spiro (cycloalkyl with 3-8 carbon atoms)) [1,3] dioxalanyl;
b is 0-6;
d is 0-6;
f = 0-6,
provided that R ¹ and R ^{2 are} not simultaneously hydrogen, and also provided that either R ¹ or R ² contains at least one (CR ³ R ⁴ ) _f OR ¹⁰ , X or cycloalkyl with 3-8 atoms carbon substituted (CR ³ R ⁴ ) _f OR ¹⁰ or a pharmaceutically acceptable salt of this compound.

 Pharmaceutically acceptable salts are those salts which are derivatives of inorganic cations such as sodium, potassium and the like; organic bases, such as mono-, di- and trialkylamines with 1-6 carbon atoms in the alkyl group, and mono-, di- and trihydroxyalkylamines with 1-6 carbon atoms in the alkyl group, and the like.

The terms “alkyl with 1-6 carbon atoms”, “alkenyl with 2-7 carbon atoms” and “alkynyl with 2-7 carbon atoms” include straight chain and branched chain compounds. Since the compounds of the present invention may contain more than one group - (CR ³ R ⁴ ) _f OR ¹⁰ , the radicals R ³ , R ⁴ , and R ¹⁰ may have the same or different meanings. Similarly, when other descriptions of substituents are repeated in the same structural formula, these substituents may be the same or different.

Аналогично, для соединения, где R¹ содержит R⁸ и R⁹, вместе образующие X, где X представляет собой 4-(2-спиро(циклоалкил с 3-8 атомами углерода))[1,3] диоксанил, циклоалкильная группа в X содержит 6 атомов углерода, a d = O,R¹ будет иметь следующую структуру:

Из числа соединений, содержащих X, предпочтительными являются те соединения, в которых алкильная группа в X, если имеется, представляет собой метильную группу, а циклоалкильная группа в X, если имеется, представляет собой циклогексил.For a compound where R ¹ contains R ⁸ and R ⁹ together forming X, where X is 5- (2,2-di (alkyl with 1-6 carbon atoms)) [1,3] dioxanyl, an alkyl group in X contains 1 carbon atom, ad = O, R ¹ will have the following structure:

Similarly, for the compound where R ¹ contains R ⁸ and R ⁹ together forming X, where X is 4- (2-spiro (cycloalkyl with 3-8 carbon atoms)) [1,3] dioxanyl, a cycloalkyl group in X contains 6 carbon atoms, ad = O, R ¹ will have the following structure:

Of the compounds containing X, those are preferred in which the alkyl group in X, if present, is a methyl group, and the cycloalkyl group in X, if present, is cyclohexyl.

When R ^{10 is} not hydrogen, alkyl, alkenyl or alkynyl, it is understood that R ¹⁰ is a group that can serve as an alcohol protecting group. So, these groups are intermediate from free hydroxylated compounds, and they are biologically active in themselves. R ¹⁰ includes three (alkyl with 1-6 carbon atoms) silyl, three (alkyl with 1-6 carbon atoms) silylethyl, triphenylmethyl, benzyl, alkoxymethyl with 2-7 carbon atoms, three (alkyl with 1-6 carbon atoms) silyl ethoxymethyl, chloroethyl and tetrahydropyranyl. Other alcohol protecting groups are also known to those skilled in the art, which are also within the scope of the present invention.

Of the compounds of the present invention, those in which R ² is hydrogen are preferred; compounds where R ² represents hydrogen, b = 0, d = 0; compounds where R ² is hydrogen, b = 0, d = 0, and each of R ⁸ and R ⁹ independently is hydrogen, alkyl or - (CR ³ R ⁴ ) _f OR ¹⁰ , or taken together form X .

Compounds of the present invention having the ester group —CO (CR ³ R ⁴ ) _b (CR ⁵ R ⁶ ) _d CR ⁷ R ⁸ R ⁹ at 42 or 31, 42 positions can be prepared by acylation of rapamycin using protected hydroxy acids or polyoxy acids alkoxycarboxylic or polyalkoxycarboxylic acids, which first activate and then, if desired, remove alcohol protecting groups from them. Several methods for activating carboxylates are known, but preferred methods are based on the use of carbodiimides, mixed anhydrides or acid chlorides. For example, a suitably substituted carboxylic acid can be activated as mixed anhydride using an acylating group such as 2,4,6-trichlorobenzoyl chloride. Treatment of rapamycin with mixed anhydride under mildly alkaline conditions provides the desired compounds. Otherwise, the acylation reaction can be carried out using 1- (3-dimethylaminopropyl) -3-ethylcarbo-diimide hydrochloride and dimethylaminopyridine. Mixtures of compounds containing esters at 42 positions and compounds containing esters at 31, 42 positions can be separated by chromatography.

The present invention also provides a process for preparing rapamycin derivatives. In particular, the process for preparing rapamycin hydroxyesters, including the compounds of the formula described above, comprises the following steps:
a) acylation of rapamycin or its functional derivative, or its analogue using an acylating agent;
or
b) sequential acylation of rapamycin or its functional derivative, or its analogue, with two acylating agents; wherein said acylating agent (or agents) is selected from acids of the formula:
HO-CO (CR ³ R ⁴ ) _b (CR ⁵ R ⁶ ) _d CR ⁷ R ⁸ R ⁹
or reactive derivatives of these compounds, where R ³ -R ⁹ , b and d have the above meanings, provided that the free hydroxyl groups are protected, and if desired, you can protect the group located in the 42 position of rapamycin or its functional derivative, with the appropriate protective groups, and after the reaction, remove any protective groups whose presence was necessary.

 The reaction can be carried out in the presence of a binder, such as a suitably substituted carbodiimide derivative. The aforementioned compounds can also be obtained by acylation using reactive derivatives of acids of formula II, such as anhydrides, mixed anhydrides or acid halides, such as chloride.

 Compounds of the invention containing an ester at 31 position and a hydroxyl group at 42 position can be obtained by protecting the alcohol group of rapamycin at 42 position with a protecting group such as a tert-butyl dimethylsilyl group, followed by esterification of the group at 31 position using the above ways. The preparation of rapamycin with a silyl ester group at the 42 position is described in US Pat. No. 5,120,842, which is hereby incorporated by reference. Removal of the protective group allows the preparation of compounds esterified at position 31. If a tert-butyl-dimethylsilyl protecting group is used, the removal of the protecting groups can be carried out under slightly acidic conditions, for example, in a mixture of acetic acid / water / tetrahydrofuran. A method for removing protecting groups is described in Example 15 of US Pat. No. 5,118,678, which is incorporated herein by reference.

 After esterifying the group at position 31 and removing the protective groups at position 42, the group at position 42 can be esterified with an acylating agent other than the acylating agent used to react with the alcohol group at position 31, to obtain compounds having different ester groups in 31 and 42 positions. In another way, compounds having esterified groups at the 42 position and prepared as described above can be reacted with another acylating agent to produce compounds having different ester groups at the 31 and 42 positions.

 The present invention also encompasses similar hydroxy acid esters obtained using other rapamycins, such as 29-demethoxy-rapamycin (US Patent 4,375,464, 32-demethoxy-rapamycin according to S.A. nomenclature); rapamycin derivatives in which double bonds at the 1-, 3- and / or 5-positions have been reduced (see US Patent 5,023,262); 29-desmethylrapamycin (see US Pat. No. 5,093,339, i.e., 32-desmethylrapamycin according to S.A. nomenclature); 7.29-bisdesmethylrapamycin (see US Pat. No. 5,093,338, 7.32-dismethylrapamycin according to S. A. nomenclature); and 15-hydroxyrapamycin (see US Patent 5102876). These patents are referred to here for information.

 The immunosuppressive activity of individual compounds of the invention was determined using a standard in vitro pharmacological test, during which the inhibition of lymphocyte proliferation (LAF test) was measured, as well as using two standard in vivo pharmacological tests. A skin graft test allows you to measure the immunosuppressive activity of the test compound, as well as the ability of the test compound to suppress or treat transplant rejection. A standard pharmacological adjuvant arthritis test measures the ability of a test compound to suppress inflammation transmitted by the immune system. The adjuvant arthritis test is a standard pharmacological test for rheumatoid arthritis. The procedures for these standard pharmacological tests are described below.

 Thymocyte proliferation induced by comitogens (i.e., LAF) was used to measure the in vitro immunosuppressive effects of representative compounds of the invention.

Thymus cells from normal BALB / C mice were cultured for 72 hours with phytohemogglutinin (PHA) and interleukin-1 and were labeled with tritium-saturated thymidine over the past six hours. Cells were cultured with and without rapamycin, cyclosporin A, or the test compound in various concentrations. Then, cells grown in culture were harvested and acquired radioactivity was determined. Inhibition of lymphoproliferation was determined as the percentage change in the counts per minute compared with control cells grown without treatment with drugs. Comparison of rapamycin data with data for each of these compounds was performed. IC _{50 was} determined for each test compound, as well as for rapamycin. Compared to representative compounds of the invention, rapamycin had an IC ₅₀ in the range of 0.6-1.5 nM. The results are given in the form of indicators IC ₅₀ and in the form of percentage inhibition of proliferation of T cells at 0.1 μm. The results obtained for representative compounds of the invention were also expressed in comparison with rapamycin data. A positive value in this ratio indicates immunosuppressive activity. A ratio of greater than 1 indicates that the test compound inhibits the proliferation of thymocytes more than rapamycin. The calculation of the indicated ratio is shown below:
IC ₅₀ rapamycin
IC ₅₀ test compound
In vivo tests were also carried out using representative compounds of the invention to determine the survival time of skin grafts taken from male BALB / c mice and transplanted into male C ₃ H (H-2K) recipients. The method used was an adapted version of the method described by Billingham RE and Medawar PBJ Exp. Biol. 28: 385-402, (1951). Briefly, a skin graft for transplantation taken from a donor was transplanted onto the back of the recipient as an allogeneic graft, and the isotransplant was used as a control in the same area. Recipients were administered various concentrations of test compounds intraperitoneally or orally. Rapamycin was used as a control. Untreated recipients were used as transplant rejection controls. The graft was observed daily and the observation results were recorded until the graft dried up and a dark scab formed. This day was considered a day of rejection. The mean graft survival (number of days ± standard deviation) in the treated group was compared with the control group. The table below shows the results. Results are expressed as average survival times per day. Graft rejection in the control group (untreated) usually occurred within 6-7 days. Test compounds were administered at a dose of 4 mg / kg.

 A standard pharmacological adjuvant arthritis test measures the ability of test compounds to prevent allergic inflammation and inhibit or treat rheumatoid arthritis. The testing method used is described below. A group of rats (male inverted Wistar Lewis rats) was previously injected with the compound to be tested (1 hour before antigen), and then Freud's complete adjuvant (FCA) was injected into the right hind paw of each rat to cause arthritis. After that, on Mondays, Wednesdays and Fridays, rats were orally administered the drug starting on day 0-14 and only 7 doses were administered. Measurements of both hind legs were carried out on days 16, 23 and 30. The difference in the volume of paws (ml) was determined from the 16th to the 0th day, then a comparison was made with the control and the percentage change in volume was derived. Inflammation in the left hind paw (where the injections were not made) was caused by inflammation transmitted by T cells; the results are noted in the above table (% change compared to control). Inflammation in the right hind paw, on the other hand, is caused by non-specific inflammation. Compounds were tested at a dose of 5 mg / kg. The results are expressed as percentage change on day 16 in the paw where the injection was not made, compared with the control; the higher the negative value of the percentage change, the more potent is the compound. Rapamycin gave values from -70% to -90% of changes in comparison with the control, and this suggests that rats that were injected with rapamycin were 70-90% less affected by allergic inflammation than rats of the control group.

 The results obtained in these standard pharmacological tests are presented after a description of the method for producing the individual compounds that have been tested. The results of these standard pharmacological tests demonstrate that the compounds of the invention have immunosuppressive activity both in vitro and in vivo.

 The results obtained in LAF tests indicate suppression of T cell proliferation, which demonstrates the immunosuppressive activity of the compounds of the invention. Further, the possibility of using the compounds of the invention as immunosuppressive agents was demonstrated by the results obtained from standard pharmacological tests on skin grafts and on adjuvant arthritis. The results of a skin graft test once again demonstrated the ability of the compounds of the invention to cure or suppress transplant rejection. The results of standard pharmacological tests for adjuvant arthritis once again demonstrated the ability of the compounds of the invention to cure or suppress rheumatoid arthritis.

 Based on the results of these standard pharmacological tests, the compounds of the invention should be considered suitable for the treatment or inhibition of transplant rejection of organs such as kidneys, heart, liver, lungs, bone marrow, pancreas (islet cells), cornea, small intestine, and skin heart valve transplants and xenografts; for the treatment or suppression of autoallergic diseases such as lupus, rheumatoid arthritis, diabetes, myasthenia gravis and multiple sclerosis; as well as inflammation such as psoriasis, dermatitis, eczema, seborrhea, intestinal inflammation, pneumonia (including asthma, chronic obstructive pulmonary disease, emphysema, acute respiratory disease syndrome, bronchitis, etc.), as well as uveitis.

 Based on the obtained activity profile, it is also believed that the compounds of the present invention have antitumor and antifungal activity, as well as antiproliferative effect. Therefore, the compounds of the invention are useful in the treatment of solid tumors, adult T-cell leukemia / lymphoma, fungal infections, and vascular hyperproliferative diseases such as restenosis and atherosclerosis. In the case of treatment of stenosis, it is desirable that the compounds of the invention are used to treat restenosis that occurs after angioplastic surgery. In this case, the compounds of the invention can be administered before the operation, during the operation, immediately after the operation, or any combination of the above administration methods can be used.

 In the treatment or inhibition of the aforementioned disease states, the compounds of the invention can be administered to mammals orally, parenterally, nasally, intrabronchially, transdermally, topically, intravaginally or rectally.

 It is contemplated that when the compounds of the invention are used as immunosuppressive or anti-inflammatory agents, they can be administered in combination with one or more other immunoregulators. Among these immunoregulators are azathioprine, corticosteroids such as prednisone and methylprednisolone, cyclophosphamide, rapamycin, cyclosporin A, FK-506, OKT-3, antithymocyte globulin (ATG). By combining the compounds of the invention with such drugs or agents in order to induce immunosuppression or to treat inflammatory processes, smaller amounts of each agent will be required to achieve the desired result. The basics of such combination therapy were developed by Stepkovsky, whose results showed that the use of a combination of rapamycin with cyclosporin A at subtherapeutic doses significantly increased the life span of a heart transplant (Transplantation Proc. 23: 507 (1991)).

 The compounds of the invention may be administered to mammals in their pure form or together with pharmaceutical carriers. The pharmaceutical carrier may be solid or liquid. It was found that for the preparation of oral compositions, a mixture of 0.01% Tween 80 in PHOSAL, PG-50 (phospholipid concentrate with 1,2-propylene glycol, manufactured by A. Nattermann & Cie. GmbH) can be used.

 A solid carrier may include substances that also act as flavorings, lubricants, diluents, suspending agents, fillers, glidants, compression aids, binders or tablet disintegrating agents; the solid carrier may be capsule material. In powder compositions, the carrier is a finely divided substance mixed with a finely divided active ingredient. In tablets, the active ingredient in appropriate proportions is mixed with a carrier that has the properties necessary for compression, and compressed into tablets of the desired shape and size. Powders and tablets preferably contain up to 99% of the active ingredient. Suitable solid carriers include, for example, calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose, methyl cellulose, sodium carboxymethyl cellulose, polyvinylpyrrolidine, low melting waxes and ion exchange resins.

 Liquid carriers are used for the preparation of solutions, suspensions, emulsions, syrups, elixirs and compositions under pressure. The active ingredient may be dissolved or suspended in a pharmaceutically acceptable liquid carrier such as water, an organic solvent, a mixture thereof or pharmaceutically acceptable oils and fats. The liquid carrier may contain other pharmaceutically acceptable additives, such as diluents, emulsifiers, buffers, preservatives, sweeteners, flavorings, suspending agents, thickeners, colorants, viscosity regulators, stabilizers or osmoregulators. Suitable examples of liquid carriers for oral and parenteral administration include water (partially containing the above additives, e.g. cellulose derivatives, preferably sodium carboxymethyl cellulose solution), alcohols (including monohydric and polyhydric alcohols, e.g. glycols) and their derivatives, as well as oils (e.g. fractionated coconut oil and peanut butter). The carrier for parenteral administration may also be an oily ester, such as ethyl oleate and isopropyl myristate. Sterile liquid carriers are used in sterile liquid compositions for parenteral administration. The liquid carrier for pressure compositions may be a halogenated hydrocarbon or other dispersant.

 Liquid pharmaceutical compositions, which are sterile solutions or suspensions, can be used, for example, for intramuscular, intraperitoneal or subcutaneous injection. Sterile solutions can also be administered intravenously.

 The compound, both in liquid and solid form, can be administered orally. The compounds of the present invention can be administered rectally in the form of conventional suppositories. For administration through the nose or as intrabronchial inhalations or insufflations, the compounds of the invention can be used in the form of aqueous or partially aqueous solutions which are used in the form of aerosols. The compounds of the invention can also be administered transdermally by applying to the skin a composition containing the active compounds and a carrier that is inert to the active compound, non-toxic to the skin and enables delivery of the absorption agent into the blood through the skin. The carrier may take a variety of forms, for example, be a cream, ointment, paste, gel and device for delivering the composition to the blood through the skin. Creams and ointments may be viscous liquids or semi-solid emulsions of either an oil-in-water type or a water-in-oil type. Also suitable are pastes consisting of absorbent powders dispersed in petrolatum or hydrophilic petrolatum containing the active ingredient. Various devices can be used to release the active ingredient into the blood, such as a semi-impermeable membrane covering the reservoir containing the active ingredient with or without a carrier, or a matrix containing the active ingredient. Other devices are described in the literature.

 In addition, the compounds of the invention can be used in the form of solutions, creams or lotions, for which formulations are prepared with pharmaceutically acceptable carriers containing 0.1-5%, preferably 2% of the active compound, which compositions are applied to areas affected by fungi.

 The dosage requirements are different for each particular composition and depend on the route of administration, the severity of the disease and the condition of the patient. Based on the results obtained in standard pharmacological tests, the estimated daily doses are 0.1 μg / kg-100 mg / kg, preferably in the range of 0.001-25 mg / kg, even better in the range of 0.001-5 mg / kg. Treatment usually begins with small doses, below the optimal dose of the compound. Gradually, the dosage is increased to achieve the optimal effect in these conditions; the exact doses for oral, parenteral, nasal, or intrabronchial administration are determined by the physician based on the experience of treating a particular patient. It is preferable to use the pharmaceutical composition in unit dose form, for example, in the form of tablets or capsules. In this form, the composition is divided into individual doses containing the desired amount of active ingredient; individual doses may be in separate packages, for example, in powders, vials, ampoules, prefilled syringes, or sachets containing liquids. One dose may be a single capsule or tablet, or you can prescribe the desired number of such capsules or tablets.

 The following examples illustrate the preparation of representative compounds of the present invention and their biological activity.

Example 1
Rapamycin 42-ester with (tetrahydropyran-2-yloxy) acetic acid
2,4,6-trichlorobenzoyl chloride (0.55 ml, 3.51 mmol) was added via syringe to a solution of glycolic acid tetrahydropyran ether (0.562 g, 3.51 mmol) and triethylamine (0.49 ml, 3.51 mmol) in 10 ml of tetrahydrofuran at 0 ^o C in nitrogen atmosphere. The mixture was stirred for 4 hours at room temperature until a white precipitate formed. The white precipitate was removed by vacuum filtration and the filtrate was concentrated in a stream of nitrogen in a warm water bath. The residue was dissolved in 10 ml of benzene, after which rapamycin (2.92 g, 3.19 mmol) and dimethylaminopyridine (0.429 g, 3.51 mmol) were added and the mixture was stirred overnight at room temperature. The mixture was diluted with EtOAc, washed with cold 1N HCl (aq), saturated NaHCO ₃ (aq) and brine, dried with MgSO ₄ , filtered and concentrated to a yellow oily solid. Flash chromatography (2X with 65% EtOAc-hexane) gave the title compound (1.114 g, 33%) as a white solid.

(-) FAB-MS) m / z 1055.5 (M ^- ), 590.3 (southern fragment), 463.2 (northern fragment).

¹ H NMR (400 MHz, d-6 dimethyl sulfoxide δ 4.60 (m, 1H, C (42) H), 4.66 (m, 1H), 4.14 (s, 2H), 3.73 (m , 1H); 3.42 (m, 1H).

¹³ C NMR (100.6 MHz, d-6 dimethyl sulfoxide δ 169.2, 97.4, 63.5, 61.2, 29.7, 24.8, 18.8.

Example 2
Rapamycin oxyacetic acid 42-ester
p-Toliolsulfonic acid (10 mg) was added to a solution of the product from Example 1 (306 mg, 0.29 mmol) in 10 ml of CH ₃ OH at 0 ^° C. The solution was stirred for 2 hours at room temperature, then the reaction was stopped with a saturated NaHCO solution ₃ . The aqueous phase was extracted with 3X EtOAc and the combined organic phases were washed with brine, dried with MgSO ₄ , filtered and concentrated to give a white solid.

(-) FAB-MS m / z 971.3 (M ^- ), 590 (southern fragment), 379.1 (northern fragment).

¹ H NMR (400 MHz, d-6 dimethyl sulfoxide) δ 4.60 (m, 1H, C (42) H), 3.98 (s, 2H).

¹³ C NMR (100.6 MHz, d-6 dimethyl sulfoxide) δ 172.1, 59.7.

The results obtained in standard pharmacological tests:
LAF IC ₅₀ : 1.80 nM
LAF Ratio: 0.83
Percentage change in adjuvant arthritis test versus control:
- 88%
Example 3
Rapamycin 42-ester with 2,2-dimethyl-3- (tetrahydropyran-2-yloxy) propionic acid
To a solution of 2,2-dimethyl-3-hydroxypropionic acid tetrahydropyran ester (0.319 g, 1.58 mmol) and triethylamine (0.22 ml, 1.58 mmol) in 5 ml of dry tetrahydrofuran at 0 ^° C under nitrogen atmosphere was added 2 , 4,6-trichlorobenzoyl chloride (0.25 ml, 1.58 mmol), the addition was carried out dropwise through a syringe. The mixture was stirred for 4.5 hours at room temperature. The white precipitate was removed by vacuum filtration and the filtrate was concentrated using a stream of nitrogen in a warm water bath. The residue was dissolved in 5 ml of benzene, then rapamycin (1.31 g, 1.43 mmol) and dimethylaminopyridine (0.193 g, 1.58 mmol) were added. The mixture was stirred overnight at room temperature, diluted with EtOAc, washed with 1N HCl (aq), saturated NaHCO ₃ (aq), H ₂ O and brine, dried with MgSO ₄ , filtered and concentrated to a yellow oily solid. Flash chromatography (1X with 60% EtOAc-hexane, 1X 55% EtOAc-hexane) gave the title compound (0.356 g, 23%) as a white solid.

(-) FAB-MS m / z 1097.7 (M ^- ), 590.4 (southern fragment).

¹ H NMR (400 MHz, d-6 dimethyl sulfoxide) δ 4.55 (m, 1H, C (42) H), 4.55 (m, 1H), 3.69 (m, 1H), 3.60 ( m, 2H), 3.42 (m, 1H), 1.13 (s, 3H), 1.11 (s, 3H).

¹³ C NMR (100.6 MHz, d-6 dimethyl sulfoxide) δ 175.0, 98.0, 73.8, 60.7, 42.6, 30.0, 24.9, 22.0, 21.6 , 18.7.

The results obtained in standard pharmacological tests:
LAF IC ₅₀ : 7.10 nM
LAP Ratio: 0.34
Example 4
Rapamycin 42-ester with 3-hydroxy-2,2-dimethylpropionic acid
p-Toluenesulfonic acid (10 mg) was added to a solution of the product from Example 3 (250 mg, 0.23 mmol) in 10 ml of CH ₃ OH at 0 ^° C. The solution was stirred for 2 hours at room temperature, then the reaction was stopped with a saturated NaHCO solution ₃ . The aqueous phase was extracted with 3X EtOAc and the combined organic phases were washed with brine, dried with MgSO ₄ , filtered and concentrated to give a white solid. Purification by flash chromatography (2X using 75% EtOAc-hexane) gave the title compound (103 mg, 45%) as a white solid.

(-) FAB-MS m / z 1013.3 (M ^- ), 590.2 (southern fragment), 421.1 (northern fragment).

¹ H NMR (400 MHz, d-6 dimethyl sulfoxide) δ 4.48 (m, 1H, C (42) H), 3.39 (d, 2H), 1.06 (s, 6H).

¹³ C NMR (100.6 MHz, d-6 dimethyl sulfoxide) δ 175.5, 68.0, 44.1, 21.7.

The results obtained in standard pharmacological tests:
LAF IC ₅₀ : 0.80 nM
LAF Ratio: 1.25
Skin graft survival time: 10.7 ± 0.5 days.

Examples 5 and 6
Rapamycin 42-ester with 2,2-dimethyl (1,3) dioxalan-4-carboxylic acid (Example 5)
31,42-diester of rapamycin with 2,2-dimethyl (1,3) dioxalan-4-carboxylic acid (Example 6)
2,4,6-trichlorobenzoyl chloride (0.56 ml, 3.61 mmol) was added via syringe to a solution of isopropylidene ketal 2,3-dioxipropionic acid (0.527 g, 3.61 mmol) and triethylamine (0.50 ml, 3.61 mmol) in 10 ml of tetrahydrofuran at 0 ^o C in a nitrogen atmosphere. The mixture was stirred for 4 hours at room temperature. The white precipitate was removed by vacuum filtration and the filtrate was concentrated using a stream of nitrogen in a warm water bath. The residue was dissolved in 15 ml of benzene, then rapamycin (3.00 g, 3.28 mmol) and dimethylaminopyridine (0.441 g, 3.61 mmol) were added and the mixture was stirred overnight at room temperature. The mixture was diluted with EtOAc, washed with cold 1N HCl (aq), saturated NaHCO ₃ (aq) and brine, dried with MgSO ₄ , filtered and concentrated to a yellow and foam. Flash chromatography on silica gel (gradient elution: 50-60-75-100% EtOAc-hexane, 4X 65% EtOAc-hexane) gave the title compound. The less polar compound with an ester group at 31, 42 positions (0.415 g) was eluted first, and the more polar compound with a monoester at 42 position (0.601 g, 16%) was eluted second, and these compounds were isolated as white solids.

Example 5
(-) FAB-MS) m / z 1041.4 (M ^- ), 590.3 (southern fragment), 449.2 (northern fragment).

¹ H NMR (400 MHz, d-6 dimethyl sulfoxide) δ 4.6 (m, 1H, C (42) H), 4.6 (m, 1H), 4.20 (dd, 1H), 3.96 ( m, 1H), 1.36 (s, 3H), 1.30 (s, 3H).

¹³ C NMR (100.6 MHz, d-6 dimethyl sulfoxide) δ 170.5, 110.2, 73.4, 66.6, 25.7, 25.4.

Example 6
(-) FAB-MS m / z 1169.6 (M ^- ).

¹ H NMR (400 MHz, d-6 dimethyl sulfoxide) δ 5.3 (m, 1H, C (31) H), 4.6 (m, 1H, C (42) H), 4.6 (m, 2H ), 4.19 (t, 1H), 4.13 (t, 1H), 3.9 (m, 2H), 1.36 (s, 3H), 1.33 (s, 3H), 1.30 (s, 3H); 1.28 (s, 3H).

¹³ C NMR (100.6 MHz, d-6 dimethyl sulfoxide) δ 170.5, 169.2, 110.3, 110.2, 73.4, 66.6, 66.5, 25.8, 25.7 , 25.4, 25.1.

The results obtained in standard pharmacological tests:
Example 5
LAF IC ₅₀ : 1.20 nM
LAF Ratio: 0.74
Example 6
LAF IC ₅₀ : 1.30 nM
LAF ratio: 0.5.

Example 7
Rapamycin 42-ester with 2,3-dioxipropionic acid
A solution of the product from Example 5 (351 mg, 0.34 mmol) in 10 ml of tetrahydrofuran and 10 ml of 1N HCl was stirred at room temperature for 6 hours. The mixture was diluted with EtOAc, washed with saturated NaHCO ₃ and brine, dried with MgSO ₄ , filtered and concentrated to an oil. Purification by flash chromatography (1X using EtOAc, 1X using 10% MeOH-CH ₂ Cl ₂ , 1X with 5% MeOH-EtOAc) gave the title compound (78 g, 23%) as a white solid.

(-) PAB-MS m / z 1001.2 (M ^- ), 590.2 (southern fragment), 409.1 (northern fragment).

¹ H NMR (400 MHz, d-6 dimethyl sulfoxide) δ 4.5 (m, 1H, C (42) H), 3.60 (m, 1H), 3.45 (m, 2H).

The results obtained in standard pharmacological tests:
LAF IC ₅₀ : 1.4 nM
LAF Ratio: 0.40
Example 8
Rapamycin 42-ester with 2,2-dimethyl (1,3) dioxane-5-carboxylic acid
2,4,6-trichlorobenzoyl chloride (0.98 ml, 6.27 mmol) was added via syringe to a solution of 2- (hydroxymethyl) -3-hydroxypropionic acid isopropylidene ketal (1,000 g, 6.24 mmol) and triethylamine (0.90 ml , 6.46 mmol) in 20 ml of tetrahydrofuran at 0 ^o C in a nitrogen atmosphere. The mixture was stirred for 4 hours at room temperature until a white precipitate formed. The white precipitate was removed by vacuum filtration and the filtrate was concentrated using a stream of nitrogen in a warm water bath. The residue was dissolved in 20 ml of benzene, then rapamycin (5.70 g, 6.24 mmol) and dimethylaminopyridine (0.762 g, 6.24 mmol) were added and the mixture was stirred overnight at room temperature. The mixture was diluted with EtOAc, washed with H ₂ O and brine, dried with MgSO ₄ , filtered and concentrated to give a yellow solid. Flash chromatography (75% EtOAc-hexane) gave the title compound (4.17 g, 63%) as a white solid.

(-) FAB-MS m / z 1055.8 (M ^- ), 590.5 (southern fragment), 463.4 (northern fragment).

¹ H NMR (400 MHz, d-6 dimethyl sulfoxide) δ 4.55 (m, 1H, C (42) H), 3.95 (m, 4H), 1.30 (s, 6H).

¹³ C NMR (100.6 MHz, d-6 dimethyl sulfoxide) δ 170.1, 97.4, 59.5, 24.8, 22.5.

The results obtained in standard pharmacological tests:
LAF IC ₅₀ : 0.76 nM
LAF Ratio: 0.45
Example 9
Rapamycin 42-ester with 3-hydroxy-2-hydroxymethylpropionic acid
A solution of the product from Example 8 (3.30 g, 3.12 mmol) in 50 ml of tetrahydrofuran and 25 ml of 1N HCI was stirred at room temperature for 2 hours. The mixture was diluted with saturated NaHCO ₃ solution and extracted with EtOAc (3X). The combined organic phases were washed with saturated NaCl (aq), dried with MgSO ₄ , filtered and concentrated to a yellow foam. Purification by flash chromatography (1X using EtOAc, 2X using 5% EtOH-EtOAc) gave the title compound (1.68 g, 53%) as a white solid.

(-) FAB-MS, m / z 1015.5 (M ^- ), 590.3 (southern fragment), 423.3 (northern fragment).

¹ H NMR (400 MHz, d-6 dimethyl sulfoxide) δ 4.6 (brs, 2H), 4.55 (m, 1H, C (42) H), 3.55 (m, 4H), 2.57- 2.53 (m, 1H).

¹³ C NMR (100.6 MHz, d-6 dimethyl sulfoxide) δ 172.2, 59.3, 51.5.

The results obtained in standard pharmacological tests:
LAF IC ₅₀ : 0.84 nM
LAF ratio: 0.57.

Example 10
Rapamycin 42-ester with 2,2,5-trimethyl (1,3) dioxane-5-carboxylic acid
Into a solution of isopropylidene ketal 2,2-bis (oxymethyl) propionic acid (1.041 g, 5.98 mmol) (prepared according to the method described by Bruice, J. Am. Chem. Soc. 89: 3568 (1967)) and triethylamine ( 0.83 ml, 5.98 mmol) in 20 ml of anhydrous tetrahydrofuran at 0 ^° C. under nitrogen atmosphere, 2,4,6-trichlorobenzoyl chloride (0.93 ml, 5.98 mmol) was added and the resulting white suspension was stirred for 5 hours at room temperature. The precipitate was removed by vacuum filtration, washing the flask and the filter cake with an additional amount (10 ml) of dry tetrahydrofuran. The filtrate was concentrated on a rotary evaporator to give a white solid. The residue was dissolved in 20 ml of dry benzene, then rapamycin (5.47 g, 5.98 mmol) and dimethylaminopyridine (0.731 g, 5.98 mmol) were added. After stirring overnight at room temperature, the mixture was diluted with EtOAc, washed with H ₂ O and saturated NaCl (aq), dried with MgSO ₄ , filtered and evaporated to give a yellow oil. Flash chromatography (5X 60% EtOAc-hexane) gave the title compound (2.2 g, 34%) as a white solid.

(-) FAB-MS m / z 1069.5 (M ^- ), 590.3 (southern fragment), 477.2 (northern fragment).

¹ H NMR (400 MHz, d-6 dimethyl sulfoxide) δ 4.57 (m, 1H, C (42) H), 4.02 (d, 2H), 3.60 (d, 2H), 1.34 ( s, 3H), 1.24 (s, 3H), 1.06 (s, 3H).

¹³ C NMR (100.6 MHz, d-6 dimethyl sulfoxide) δ 173.2, 99.0, 65.0, 22.2, 18.1.

The results obtained in standard pharmacological tests:
LAF IC ₅₀ : 4.90 nM
LAF Ratio: 0.41
Skin graft survival: 11.0 ± 1.3 days.

Example 11
Rapamycin 42-ester with 2,2-bis- (hydroxymethyl) propionic acid
A solution of the product from Example 10 (2.8 g, 2.65 mmol) in 50 ml of tetrahydrofuran and 25 ml of 1N HCl was stirred at room temperature for 4 hours. The mixture was diluted with water and extracted three times with EtOAc. The combined organic phases were washed with saturated NaHCO ₃ , saturated NaCl, dried with MgSO ₄ , filtered and evaporated to give a yellow oily solid. Purification by flash chromatography (3X using EtOAc) gave the title compound (1.6 g, 59%).

(-) FAB-MS m / z 1029.6 (M ^- ), 590.4 (southern fragment), 437.3 (northern fragment).

¹ H NMR (400 MHz, d-6 dimethyl sulfoxide) δ 4.5 (m, 1H, C (42) H), 3.45 (s, 4H), 1.04 (s, 3H).

¹³ C NMR (100.6 MHz, d-6 dimethyl sulfoxide) δ 174.2, 63.7, 63.6, 49.9, 16.8.

The results obtained in standard pharmacological tests:
LAF IC ₅₀ : 0.80 and 1.80 nM
LAF ratio: 1.00 and 0.44
Skin graft survival: 11.4 ± 1.5 and 12.0 ± 1.1 days.

 Percentage change in adjuvant arthritis compared to control: -88%.

Example 12
Rapamycin 42-ester with 2,2-dimethyl-5- (2-trimethylsilyanylethoxymethyl) (1,3) dioxane-5-carboxylic acid
2,4,6-trichlorobenzoyl chloride (0.14 ml, 0.86 mmol) was added via syringe to the isopropylidene ketal solution of 2,2-bis- (hydroxymethyl) -2- (2-trimethylsilylethoxy) propionic acid (0.250 g, 0.86 mmol) and triethylamine (0.12 ml, 0.86 mmol) in 2 ml of tetrahydrofuran at 0 ^o C in a nitrogen atmosphere. The mixture was stirred for 4 hours at room temperature until a white precipitate formed. The white precipitate was removed by vacuum filtration, the filtrate was concentrated using a stream of nitrogen in a warm water bath. The residue was dissolved in 20 ml of benzene, then rapamycin (0.786 g, 0.86 mmol) was added and the mixture was stirred overnight at room temperature. The mixture was diluted with EtOAc, washed with H ₂ O and brine, dried with MgSO ₄ filtered and concentrated to give a yellow solid. Flash chromatography (elution gradient: 40-60-80-100% EtOAc-hexane) gave the title compound (0.559 g, 54%) as a white solid.

(-) FAB-MS m / z 1185.2 (M ^- ), 590.1 (southern fragment), 593 (northern fragment).

¹ H NMR (400 MHz, d-6 dimethyl sulfoxide] δ 4.55 (m, 1H, C (42) H), 3.73 (m, 4H), 3.57 (s, 2H), 3.43 ( t, 2H), 1.29 (s, 6H), 0.79 (t, 2H), -0.04 (s, 9H).

¹³ C NMR (100.6 MHz, d-6 dimethyl sulfoxide) δ 171.1, 97.7, 70.2, 68.1, 61.3, 46.0, 24.6, 22.1, 14.6 , -1.3.

The results obtained in standard pharmacological tests:
LAF IC ₅₀ : 7.20 nM
LAF ratio: 0.05.

Examples 13 and 14
Rapamycin 42-ester with 3-methyl-1,5-dioxaspiro (5.5) undecane-3-carboxylic acid (Example 13) and
31,42-diester of rapamycin with 3-methyl-1,5-dioxa-spiro (5.5) undecane-3-carboxylic acid (Example 14)
2,4,6-trichlorobenzoyl chloride (0.16 ml, 1.0 mmol) was added via syringe to a solution of 2,3-dioxipropionic acid cyclohexylidene ketal (0.214 g, 1.0 mmol) and triethylamine (0.14 ml, 1.0 mmol) in 2.5 ml of tetrahydrofuran at 0 ^o C in a nitrogen atmosphere. The mixture was stirred for 4 hours at room temperature. The white precipitate was removed by vacuum filtration, the filtrate was concentrated using a stream of nitrogen in a warm water bath. The residue was dissolved in 3 ml of benzene, then rapamycin (0.457 g, 0.5 mmol) and dimethylaminopyridine (0.061 g, 0.5 mmol) were added and the mixture was stirred overnight at room temperature. The mixture was diluted with EtOAc, cold IN HCl solution (waves), saturated NaHCO ₃ solution (aq), and brine, dried with MgSO ₄ , filtered and concentrated to a yellow foam. Flash chromatography (45-50% EtOAc-hexane) gave the title compound. A compound with an ester group at 31, 42 positions (0.168 g, 26%) was eluted first, and a more polar compound with a monoester group at 42 positions (0.301 g, 52%) was eluted second, and products were isolated as white solids.

Example 13
(-) FAB-MS m / z 1109.5 (M ^- ), 590.3 (southern fragment), 517.3 (northern fragment).

¹ H NMR (400 MHz, d-6 dimethyl sulfoxide) δ 4.55 (m, 1H, C (42) H), 3.61 (t, 4H), 1.04 (s, 3H).

¹³ C NMR (100.6 MHz, d-6 dimethyl sulfoxide) δ 173.3, 97.2, 64.2.

Example 14
(-) FAB-MS m / z 1305.6 (M ^- ).

¹ H NMR (400 MHz, d-6 dimethyl sulfoxide) δ 5.25 (m, 1H, C (31) H), 4.55 (m, 1H, C (42) H), 3.64-3.54 (m, 8H); 1.05 (s, 3H); 0.97 (s, 3H).

¹³ C NMR (100.6 MHz, d-6 dimethyl sulfoxide) δ 173.2, 172.1, 97.3, 97.2, 64.3, 64.2, 63.9.

The results obtained in standard pharmacological tests:
Example 13
LAF IC ₅₀ : 0.6 nM
LAF ratio: 2.00.

Example 14
LAF: inhibited T cell proliferation by 43% at 0.1 μm.

Claims (21)

1. Hydroxyesters of rapamycin of General formula I

where each of R ¹ and R ² independently represent hydrogen or —CO (CR ³ R ⁴ ) _b (CR ⁵ R ⁶ ) _d CR ⁷ R ⁸ R ⁹ ;
each of R ³ and R ⁴ independently represents hydrogen, alkyl with 1-6 carbon atoms;
each of R ⁵ and R ⁶ independently represents hydrogen, alkyl with 1-6 carbon atoms, or R ⁵ and R ⁶ can together form a group X or a cycloalkyl ring with 3-8 carbon atoms;
R ⁷ represents hydrogen, alkyl with 1-6 carbon atoms, (CR ³ R ⁴ ) _f OR ¹⁰ ;
each of R ⁸ and R ⁹ independently represents hydrogen, alkyl with 1-6 carbon atoms, - (CR ³ R ⁴ ) _f OR ¹⁰ or R ⁸ and R ⁹ can together form a group X or a cycloalkyl ring with 3-8 carbon atoms;
R ¹⁰ represents hydrogen, alkyl with 1-6 carbon atoms, three (alkyl with 1-6 carbon atoms) silyl, three (alkyl with 1-6 carbon atoms) silylethyl, or tetrahydropyranyl;
X represents 5- (2,2-di (alkyl with 1-6 carbon atoms)) (1,3) dioxanyl, 5- (2-spiro (cycloalkyl with 3-8 carbon atoms)) (1,3) dioxanyl 4- (2,2-di (alkyl with 1-6 carbon atoms)) (1,3) -dioxanyl, 4- (2-spiro (cycloalkyl with 3-8 carbon atoms)) (1,3) dioxanyl;
b is 0-6;
d is 0-6;
f = 0-6,
provided that R ¹ and R ^{2 are} not simultaneously hydrogen, and also provided that either R ¹ or R ² contains at least one (CR ³ R ⁴ ) _f OR ¹⁰ , X or cycloalkyl with 3-8 atoms carbon, or a pharmaceutically acceptable salt of this compound. 2. The compound according to claim 1, where R ² represents hydrogen, or their pharmaceutically acceptable salt of this compound.  3. The compound according to claim 2, where b = 0 and d = 0, or a pharmaceutically acceptable salt of this compound. 4. The compound according to claim 3, where each of R ⁸ and R ⁹ independently represent hydrogen, alkyl, or - (CR ³ R ⁴ ) _f OR ¹⁰ , or together form X, or a pharmaceutically acceptable salt of this compound.  5. The compound according to claim 1, which is rapamycin having a 42 position ether (tetrahydropyran-2-yloxy) acetic acid ester or a pharmaceutically acceptable salt of this compound.  6. The compound according to claim 1, which is a rapamycin having an oxyacetic acid ester at the 42 position, or a pharmaceutically acceptable salt of this compound.  7. The compound according to claim 1, which is a rapamycin 42-ester of 2,2-dimethyl-3- (tetrahydropyran-2-yloxy) propionic acid, or a pharmaceutically acceptable salt of this compound.  8. The compound according to claim 1, which is a rapamycin 42-ester with 3-hydroxy-2,2-dimethyl-propionic acid, or a pharmaceutically acceptable salt of this compound.  9. The compound according to claim 1, which is a rapamycin 42-ester with 2,2-dimethyl- (1,3) dioxalan-4-carboxylic acid, or a pharmaceutically acceptable salt of this compound.  10. The compound according to claim 1, which is a 31.41-diester of rapamycin with 2,2-dimethyl- (1,3) dioxalan-4-carboxylic acid, or a pharmaceutically acceptable salt of this compound.  11. The compound according to claim 1, which is a rapamycin 42-ester with 2,3-dihydroxypropionic acid, or a pharmaceutically acceptable salt of this compound.  12. The compound according to claim 1, which is a rapamycin 42-ester with 2,2-dimethyl- (1,3) dioxane-5-carboxylic acid, or a pharmaceutically acceptable salt of this compound.  13. The compound according to claim 1, which is a rapamycin 42-ester with 3-hydroxy-2-hydroxymethylpropionic acid, or a pharmaceutically acceptable salt of this compound.  14. The compound according to claim 1, which is a rapamycin 42-ester of 2,2,5-trimethyl- (1,3) dioxane-5-carboxylic acid, or a pharmaceutically acceptable salt of this compound.  15. The compound according to claim 1, which is a 42-ester of rapamycin with 2,2-bis- (hydroxymethyl) propionic acid, or a pharmaceutically acceptable salt of this compound.  16. The compound according to claim 1, which is a rapamycin 42-ester with 2,2-dimethyl-5- (2-trimethylsilanylethoxymethyl) (1,3) dioxane-5-carboxylic acid, or a pharmaceutically acceptable salt of this compound.  17. The compound according to claim 1, which is a rapamycin 42-ester with 3-methyl-1,5-dioxa-spiro (5,5) undecane-3-carboxylic acid, or a pharmaceutically acceptable salt of this compound.  18. The compound according to claim 1, which is a 31, 42-rapamycin diester with 3-methyl-1,5-dioxa-spiro (5,5) undecane-3-carboxylic acid, or a pharmaceutically acceptable salt of this compound.  19. A method of treating a condition associated with an incompatibility reaction during transplantation in mammals in need of such treatment, comprising administering to said mammal an effective amount of a compound of formula I according to claim 1 or a pharmaceutically acceptable salt of this compound.  20. A method of treating rheumatoid arthritis in mammals in need of such treatment, which comprises administering to said mammal an effective amount of a compound of formula I according to claim 1 or a pharmaceutically acceptable salt of this compound. 21. The method of producing rapamycin hydroxyesters of the general formula I according to claim 1, wherein the rapamycin or its functional derivatives are reacted with an acylating agent of the formula II
HO-CO (CR ³ R ⁴ ) _b (CR ⁵ R ⁶ ) _d CR ⁷ R ⁸ R ⁹ ,
in which R ³ - R ⁹ , b and d have the meanings given in claim 1,
provided that the free hydroxyl groups are protected and, if desired, the group located in the 42 position of rapamycin or its functional derivative is protected with the appropriate protective group, followed by acylation, removing any protective groups whose presence was necessary.